番茄红素对人骨肉瘤细胞生长的影响及机制研究

Effect of lycopene on the growth of human osteosarcoma cells and its mechanism

目的研究番茄红素(Lycopene,LP)对人骨肉瘤MG-63细胞增殖和凋亡的影响并初步探讨其机制。方法取处于对数生长期的人骨肉瘤MG-63细胞,设空白对照组,LP低、中、高剂量组(5、10、20μg/m L)和顺铂(40μg/m L)组;给药干预48 h,观察细胞生长状况,MTT比色法检测细胞增殖抑制率,流式细胞术(Flow cytometry,FCM)检测细胞周期、细胞凋亡状况并计算凋亡率,逆转录PCR(RTPCR)法检测凋亡相关基因Bax mRNA、bcl-2 mRNA表达并计算Bax/bcl-2值,免疫蛋白印迹法(Western blot)检测凋亡相关蛋白caspase-3表达。结果光学显微镜下观察可见,空白对照组人骨肉瘤MG-63细胞呈贴壁生长,增殖迅速,而LP组细胞增殖受到抑制,细胞数明显减少,呈现回缩、脱落现象。与空白对照组比较发现经LP干预48 h能够显著提高人骨肉瘤MG-63细胞增殖抑制率,显著延长细胞周期G_0/G_1期并缩短S期、G_2/M期,显著提高细胞凋亡率(Apoptosis index,AI),上调Bax mRNA表达并下调bcl-2 mRNA表达,显著提高Bax/bcl-2比值,显著上调caspase-3蛋白表达,差异均具有统计学意义(P<0.05或P<0.01)。结论 LP具有抑制人骨肉瘤细胞增殖并促进其凋亡的药理学作用,机制可能与阻滞细胞周期和影响凋亡调控基因、蛋白表达有关。

Objective The objective of this study was to investigate the effect of lycopene（ LP） on the proliferation and apoptosis of human osteosarcoma MG-63 cells and to explore its mechanism. Methods MG-63 cells in the logarithmic growth phase were treated with LP（ 5,10 and 20 μg/m L） or Cisplatin（ 40 μg/m L） for48 h. No LP or Ciplatin treatment was as the control group. The cell growth status was observed and the cell proliferation was measured by MTT colorimetric assay. Flow cytometry（ FCM） was used to detect the cell cycle and apoptosis of MG-63 cells and the apoptotic rate was calculated in this study. Reverse transcription-polymerase chain reaction（ RT-PCR） was used to measure mRNA levels of Bax and bcl-2,and calculate the ratio of Bax/bcl-2 in MG-63 cells. Western blot also was used to examine the expression of caspase-3 protein in MG-63 cells. Results MG-63 cells were adherent and proliferated rapidly in the control group. The cells in LP treated groups showed that the cell proliferation was inhibited and the number of cells was significantly decreased,with retraction and detaching. The inhibitory rate of MG-63 cells in LP treated group were significantly increased when compared to the control group. LP significantly prolonged the cell cycle at the G_0/G_1 phase and shortened the cell cycle at the S and G_2/M phase. LP also significantly increased the apoptosis rate,up-regulated the mRNA level of Bax,down-regulated the mRNA level of bcl-2,increased the ratio of Bax/bcl-2 and the expression of caspase-3 protein in MG-63 cells（ P 0. 05 or P 0. 01）. Conclusion LP may effectively inhibit the proliferation of MG-63 cells and promote the apoptosis. The mechanism may be related to the cell cycle and the regulation of apoptosis and gene expression.